Vincent Laurent

Inactivation of the infralimbic but not the prelimbic cortex impairs consolidation and retrieval of fear extinction

Rats were subjected to one or two cycles of context fear conditioning and extinction to study the roles of the prelimbic cortex (PL) and infralimbic cortex (IL) in learning and relearning to inhibit fear responses. Inactivation of the PL depressed fear responses across the first or second extinction but did not impair learning or relearning fear inhibition (experiment 1). Inactivation of the IL did not affect inhibition across the first extinction but disrupted its long-term retention. Inactivation of the IL impaired inhibition across the second extinction, and inactivation before or after this extinction impaired long-term retention (experiments 2 and 3). Inactivation of the IL before the retention test restored extinguished fear responses (experiment 4). These results show for the first time that neuronal activity in the PL is involved in the expression of fear responses but not in the learning that underlies long-term fear inhibition. They also confirm that the IL is involved in this inhibitory learning: Specifically, they show that the IL is critical for consolidation and retrieval of this inhibitory learning. The role of the IL is discussed in terms of a contemporary neural model of fear extinction.

Distinct contributions of the basolateral amygdala and the medial prefrontal cortex to learning and relearning extinction of context conditioned fear

We studied the roles of the basolateral amygdala (BLA) and the medial prefrontal cortex (mPFC) in learning and relearning to inhibit context conditioned fear (freezing) in extinction. In Experiment 1, pre-extinction BLA infusion of the NMDA receptor (NMDAr) antagonist, ifenprodil, impaired the development and retention of inhibition but post-extinction infusion spared retention. Pre-extinction infusion of the GABA(A) agonist, muscimol, depressed freezing and impaired retention as did post-extinction infusion. In Experiment 2, pre-extinction mPFC infusion of ifenprodil spared the development of inhibition whereas muscimol depressed freezing. Both impaired retention when infused pre- or post-extinction. Thus, the development of inhibition involves NMDAr activation in the BLA, whereas its consolidation involves both NMDAr activation in the mPFC and NMDAr-independent mechanisms in the BLA. In Experiment 3, BLA infusion of ifenprodil impaired relearning and retention of inhibition when infused before but did not impair retention when infused after re-extinction. BLA infusion of muscimol depressed freezing but did not impair retention when infused before or after re-extinction. In Experiment 4, mPFC infusion of ifenprodil impaired relearning when infused before re-extinction, whereas muscimol depressed responses. Both drugs impaired retention when infused into the mPFC before or after re-extinction. Thus, relearning to inhibit fear responses involves NMDAr activation in both the BLA and mPFC and consolidation of the inhibitory memory involves NMDAr activation in the mPFC. However, relearning and consolidation occur in the absence of neuronal activity within the BLA. We propose that NMDAr in the mPFC supports relearning inhibition when the BLA is inactivated.

The basolateral amygdala is necessary for learning but not relearning extinction of context conditioned fear

Extinction of conditioned fear involves new learning that inhibits but does not eliminate the original fear memory. This inhibitory learning is thought to require activation of NMDA receptors (NMDAr) within the basolateral amygdala (BLA). However, once extinction has been learned, the role played by the BLA during subsequent extinction procedures remains unknown. The present study examined the role of neuronal activity and NMDAr activation in rats receiving their first or second extinction of context fear. We found that BLA infusion of DL-APV, a competitive antagonist of NMDAr, depressed fear responses at both the first and second extinction. It impaired learning extinction but spared and even facilitated relearning extinction. BLA infusion of muscimol, a GABA(A) agonist, produced a similar outcome, suggesting that DL-APV not only blocked NMDAr-dependent plasticity but also disrupted neuronal activity. In contrast, infusion of ifenprodil, a more selective antagonist of NMDAr containing the NR2B subunit, did not depress fear responses but impaired short- and long-term inhibition of fear at both the first and second extinction. Therefore, we suggest that relearning extinction normally requires NMDAr containing the NR2B subunit in the BLA. However, simultaneous blockade of these receptors and neuronal activity in the BLA results in compensatory learning that is able to promote long-term re-extinction. These data are consistent with a current model that attributes fear extinction to interactions between several neural substrates, including the amygdala and the medial prefrontal cortex.

μ- and δ-Opioid-Related Processes in the Accumbens Core and Shell Differentially Mediate the Influence of Reward-Guided and Stimulus-Guided Decisions on Choice

Two motivational processes affect choice between actions: (1) changes in the reward value of the goal or outcome of an action and (2) changes in the predicted value of an action based on outcome-related stimuli. Here, we evaluated the role of μ-opioid receptor (MOR) and δ-opioid receptor (DOR) in the nucleus accumbens in the way these motivational processes influence choice using outcome revaluation and pavlovian-instrumental transfer tests. We first examined the effect of genetic deletion of MOR and DOR in specific knock-out mice. We then assessed the effect of infusing the MOR antagonist d-Phe-Cys-Tyr-d-Trp-Arg-Thr-Pen-Thr-NH2 (CTAP) or the DOR antagonist naltrindole into the core or shell subregions of the nucleus accumbens on these tests in rats. We found that, whereas MOR knock-outs showed normal transfer, they failed to show a selective outcome revaluation effect. Conversely, DOR knock-outs showed normal revaluation but were insensitive to the influence of outcome-related cues on choice. This double dissociation was also found regionally within the nucleus accumbens in rats. Infusion of naltrindole into the accumbens shell abolished transfer but had no effect on outcome revaluation and did not influence either effect when infused into the accumbens core. Conversely, infusion of CTAP into the accumbens core abolished sensitivity to outcome revaluation but had no effect on transfer and did not influence either effect when infused into the accumbens shell. These results suggest that reward-based and stimulus-based values exert distinct motivational influences on choice that can be doubly dissociated both neuroanatomically and neurochemically at the level of the nucleus accumbens.

Blockade of Dopamine Activity in the Nucleus Accumbens Impairs Learning Extinction of Conditioned Fear.

Three experiments used rats to investigate the role of dopamine activity in learning to inhibit conditioned fear responses (freezing) in extinction. In Experiment 1, rats systemically injected with the D2 dopamine antagonist, haloperidol, froze more across multiple extinction sessions and on a drug-free retention test than control rats. In Experiment 2, rats extinguished under an intracerebroventricular (ICV) infusion of haloperidol suppressed fear responses across extinction but froze more on a subsequent drug-free retention test than control rats. In Experiment 3, rats extinguished under an infusion of haloperidol in the nucleus accumbens were impaired in suppressing fear responses across extinction and froze more on subsequent drug-free retention test than control rats. These results show that learning to inhibit fear responses in extinction requires dopamine activity in the nucleus accumbens. They were interpreted to mean that dopaminergic activity in the nucleus accumbens regulates the prediction error required for learning to inhibit fear responses in extinction.

Subchronic phencyclidine treatment impairs performance of C57BL/6 mice in the attentional set-shifting task.

Schizophrenic patients show deficits in their ability to shift attention to new stimuli during attentional set-shifting tasks. An equivalent task has been developed in rodents. This task was used to assess effects of phencyclidine (PCP), a drug widely used to model schizophrenia, in mice. Mice were administered PCP (0.63 or 1.3 mg/kg) daily, beginning 5 days prior to training and continuing throughout the test period. Mice were trained to discriminate stimuli to find a food reward in a series of discriminations. PCP increased the number of trials required to reach the criterion during simple and reversal discriminations, but not when the discriminations were repeated with a new set of stimuli (i.e. during the intradimensional shift and its reversal), suggesting an interaction between PCP effects and novelty. The 1.3 mg/kg of PCP also impaired performance of mice during the extradimensional shift. Similar results have been reported in schizophrenic patients. Although, in control mice, performance of the extradimensional shift was not impaired relative to the intradimensional shift, suggesting that the mouse may not be a suitable species for assessment in this model, specific discriminations within this task were sensitive to impairment by PCP, implying, therefore, that it may have some utility in modelling discrete aspects of schizophrenia in humans.

Learning-Related Translocation of δ-Opioid Receptors on Ventral Striatal Cholinergic Interneurons Mediates Choice between Goal-Directed Actions

The ability of animals to extract predictive information from the environment to inform their future actions is a critical component of decision-making. This phenomenon is studied in the laboratory using the pavlovian–instrumental transfer protocol in which a stimulus predicting a specific pavlovian outcome biases choice toward those actions earning the predicted outcome. It is well established that this transfer effect is mediated by corticolimbic afferents on the nucleus accumbens shell (NAc-S), and recent evidence suggests that δ-opioid receptors (DORs) play an essential role in this effect. In DOR-eGFP knock-in mice, we show a persistent, learning-related plasticity in the translocation of DORs to the somatic plasma membrane of cholinergic interneurons (CINs) in the NAc-S during the encoding of the specific stimulus–outcome associations essential for pavlovian–instrumental transfer. We found that increased membrane DOR expression reflected both stimulus-based predictions of reward and the degree to which these stimuli biased choice during the pavlovian–instrumental transfer test. Furthermore, this plasticity altered the firing pattern of CINs increasing the variance of action potential activity, an effect that was exaggerated by DOR stimulation. The relationship between the induction of membrane DOR expression in CINs and both pavlovian conditioning and pavlovian–instrumental transfer provides a highly specific function for DOR-related modulation in the NAc-S, and it is consistent with an emerging role for striatal CIN activity in the processing of predictive information. Therefore, our results reveal evidence of a long-term, experience-dependent plasticity in opioid receptor expression on striatal modulatory interneurons critical for the cognitive control of action.

δ-Opioid and Dopaminergic Processes in Accumbens Shell Modulate the Cholinergic Control of Predictive Learning and Choice

Decision-making depends on the ability to extract predictive information from the environment to guide future actions. Outcome-specific Pavlovian-instrumental transfer (PIT) provides an animal model of this process in which a stimulus predicting a particular outcome biases choice toward actions earning that outcome. Recent evidence suggests that cellular adaptations of δ-opioid receptors (DORs) on cholinergic interneurons (CINs) in the nucleus accumbens shell (NAc-S) are necessary for PIT. Here we found that modulation of DORs in CINs critically influences D1-receptor (D1R)-expressing projection neurons in the NAc-S to promote PIT. First, we assessed PIT-induced changes in signaling processes in dopamine D1- and D2-receptor-expressing neurons using drd2-eGFP mice, and found that PIT-related signaling was restricted to non-D2R-eGFP-expressing neurons, suggesting major involvement of D1R-neurons. Next we confirmed the role of D1Rs pharmacologically: the D1R antagonist SCH-23390, but not the D2R antagonist raclopride, infused into the NAc-S abolished PIT in rats, an effect that depended on DOR activity. Moreover, asymmetrical infusion of SCH-23390 and the DOR antagonist naltrindole into the NAc-S also abolished PIT. DOR agonists were found to sensitize the firing responses of CINs in brain slices prepared immediately after the PIT test. We confirmed the opioid-acetylcholinergic influence over D1R-neurons by selectively blocking muscarinic M4 receptors in the NAc-S, which tightly regulate the activity of D1Rs, a treatment that rescued the deficit in PIT induced by naltrindole. We describe a model of NAc-S function in which DORs modulate CINs to influence both D1R-neurons and stimulus-guided choice between goal-directed actions.

Role of the basolateral amygdala in the reinstatement and extinction of fear responses to a previously extinguished conditioned stimulus.

Four experiments used rats to study the role of the basolateral amygdala (BLA) in the reinstatement and extinction of fear responses (freezing) to a previously extinguished conditioned stimulus (CS). In Experiment 1, BLA inactivation before pairing the extinguished CS with the shock unconditioned stimulus (US) or before US-alone exposure impaired the restoration and the reinstatement of fear responses to the extinguished CS. In Experiment 2, BLA inactivation before extinction impaired long-term inhibition of fear responses, but its inactivation before extinction of fear responses restored by CS-US pairing did not impair long-term inhibition. In Experiment 3, BLA inactivation before extinction of fear responses or before the extinction of fear responses reinstated by US-alone exposure impaired long-term inhibition. In Experiment 4, BLA inactivation did not impair long-term inhibition of fear responses reinstated by US-alone exposure if the context where the US-alone exposure occurred had been previously extinguished. These results confirm that the BLA is critical for both learning fear and fear inhibition, but not for relearning this inhibition. The results are consistent with the view that reinstatement is due to the extinguished CS being tested in a dangerous context and are discussed in terms of a contemporary neural model of fear inhibition.

Infusion of the NMDA receptor antagonist, DL-APV, into the basolateral amygdala disrupts learning to fear a novel and a familiar context as well as relearning to fear an extinguished context

Ample evidence suggests that activation of NMDA receptors (NMDAr) in the basolateral complex of the amygdala (BLA) is necessary for context fear conditioning. The present series of experiments examined whether this activation was still required when the to-be-shocked context had a history. We found that BLA infusion of the selective NMDAr antagonist DL-APV impaired the acquisition of fear responses to a novel context, a moderately familiar context, or a highly familiar context. The same treatment also impaired the reacquisition of fear responses to a dangerous context, a context extinguished to criterion, or a context massively extinguished. Importantly, DL-APV persistently suppressed fear responses, suggesting that the NMDAr antagonist disrupted basal synaptic transmission in the BLA. Therefore, we conclude that neuronal activity in the BLA is critical for learning and relearning context-conditioned fear. This finding is consistent with current neural models that attribute context fear conditioning to interactions among several brain regions, most notably the hippocampus and the BLA.